Optimization of the under-agarose assay of porcine neutrophil migration.
Important procedural factors in the under-agarose assay for porcine neutrophil migration were identified, and optimal conditions were established. Three factors were tested: the concentration of zymosan-activated serum inoculated into the outer well; the number of neutrophils inoculated into the center well; and the time of incubation of the agarose plates. All factors had a significant (P less than 0.0001, 0.0001, and 0.01, respectively) effect on the chemotactic index of porcine neutrophils. The optimal combination of these 3 factors was undiluted zymosan-activated serum as the chemoattractant, 8 X 10(5) neutrophils inoculated into the center well, and 5 hours of incubation. The assay was validated, using standard conditions, and the data were used to predict the number of pigs and/or repetitive assays needed to identify differences among experimental groups.
Elliott
G S
GS
Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47906.
Rebar
A H
AH
McCabe
G P
GP
Alzola
C
C
D000818:Animals; D002464:Cell Migration Inhibition; D002499:Centrifugation, Density Gradient; D002634:Chemotaxis, Leukocyte; D005362:Ficoll; D009504:Neutrophils; D012685:Sepharose; D013552:Swine / Q000097:blood*; D015054:Zymosan / Q000097:blood
D005362:Ficoll; D015054:Zymosan; D012685:Sepharose